Comparison of effects of dietary‐specific fatty acids on growth and lipid metabolism in Nile tilapia

The dominant fatty acids (FAs) in oils are often used to explain different nutritional effects of dietary oils in fish. However, the amounts of dominant FAs among oils are different, and the nutritional roles of these important FAs in fish have not been precisely compared at similar levels in feeding trials. In the present study, different amounts of palmitic acid were added to safflower oil (SO), olive oil (OO) and fish oil (FO) to obtain comparable amounts (about 550 g/kg of total FAs) of 18:2n‐6, 18:1n‐9 and 20:5n‐3 + 22:6n‐3 and subsequently fed to Nile tilapia (11.1 ± 0.01 g) for 8 weeks. The results showed similar growth among groups but FO group obtained lower fat deposition, serum ALT and AST activities, compared to OO. Lipogenesis‐related gene expressions were higher in OO group than FO group in liver, muscle and adipose tissue, but there were only few differences in these genes between SO and FO groups. Lipid catabolism genes in FO group were higher than OO and SO groups in adipose tissue, but not in muscle, and the significantly higher expressions of CPT1b and PPARα were only observed in liver. Overall, dietary 18:2n‐6, 20:5n‐3 and 22:6n‐3 were beneficial to normal growth and lipid metabolism, whereas high amount of 18:1n‐9 induced lipid deposition and liver damage in Nile tilapia.     

基于侯选基因标记的四倍体马铃薯休眠QTL关联分析

休眠期是马铃薯(SolanumtuberosumL.)重要的块茎性状之一,寻找调控马铃薯块茎休眠的关键基因,揭示其分子机制以选育具有适宜休眠期长度的马铃薯品种,对于解决当前马铃薯产业中过长或过短休眠期带来的经济损失和食品安全隐患等问题十分关键。前期研究在二倍体马铃薯连锁群体中定位了6个加性休眠QTL,本研究拟在四倍体马铃薯育种材料中验证这些休眠QTL。基于休眠QTL连锁的候选基因标记,采用混合线性模型(MLM),模型中考虑群体结构和亲缘关系(Q+K),在四倍体马铃薯自然群体St-hzau中对马铃薯块茎休眠期进行了关联分析。5号染色体上休眠QTL DorB5.3连锁的候选基因标记S199_300和GWD (根据葡聚糖水双激酶α-glucan water dikinase基因设计)与马铃薯块茎休眠期具有显著的关联(P0.05),分别解释了休眠期表型变异的7.8%和3.2%,分别能增加休眠期7.1 d和4.5 d,即在二倍体马铃薯连锁群体中定位的稳定主效休眠QTL DorB5.3在四倍体马铃薯关联群体St-hzau中也表现显著, DorB5.3的稳定性在关联分析结果中得到了验证,表明候选基因标记策略在马铃薯块茎休眠QTL关联分析中是一种有效的策略。本研究所验证的主效休眠QTL DorB5.3及相应连锁标记可以直接用于马铃薯休眠育种。据此可以推测GWD可能在控制还原糖含量和块茎休眠2个方面均发挥作用,马铃薯块茎休眠机制与还原糖含量变化机制可能存在着部分交叉。     

基于高光谱和参数优化支持向量机的水稻施氮水平分类研究

为探索水稻氮素营养的快速、无损诊断方法以及构建基于高光谱技术的水稻氮素营养状况分类识别模型。本研究以 4种不同施氮水平的“中嘉早 17”水稻分蘖期顶部第三完全展开叶叶片(简称顶三叶)为研究对象，测定各叶片的可见光到近红外波段(350～ 2500 nm)内的光谱数据，对所获取的光谱数据进行平滑处理和归一化处理，以消除噪声及量纲的影响，并采用主成分分析(PCA)的方法进行数据降维至 22维，同时分别选用基于网格搜索算法、粒子群算法和遗传算法优化参数的支持向量机进行水稻氮素营养状况分类识别模型的建立。研究结果表明:1)不同施氮水平下的水稻叶片光谱反射率曲线走势大致相同，但不同施氮水平下 780～ 1 300、1 400～ 1 850及 1 900～ 2500 nm波段光谱反射率存在一定的差别；2)优化参数后的 SVM模型与默认参数下的 SVM模型相比，其训练集与测试集分类识别效果都要优于默认参数下的 SVM模型。其中，以遗传算法优化参数的 SVM模型识别分类效果最佳，训练集和测试集识别准确率分别为 99.375%、98.750%，测试集的4种施氮水平(施氮量从低到高)识别准确率分别为 100%、95%、100%和 100%。结果表明利用高光谱技术能够很好地进行水稻氮素营养状况的定性诊断研究。为快速水稻氮素营养诊断提供了一种新途径，为精确施氮提供了技术支撑和理论依据。     

生长后期干旱复水对饲草大麦产量、品质及叶绿素含量的影响

为研究水分胁迫对饲草大麦生长、产量、籽粒关键品质和相关生理指标的影响,于2017～2018年采用盆栽称重控水法对甘饲麦1号进行以干旱胁迫持续时间(T)为主因素(10、20、30 d)和旱后复水量为副因素(W1、W2和W3)的裂区试验,测定其株高、穗长、生育期、单株产量及构成因子、关键品质指标以及叶绿素含量的变化。结果显示:干旱胁迫时间显著影响了各项测定指标,各项测定指标均表现出随着干旱胁迫时间的延长其显著降低,指标测定值均表现出T1T2T3。较T1,T2和T3处理的株高显著降低了10.87和16.26 cm,生育期显著提前了5.12和14.00 d;穗粒数、千粒重和单株产量分别显著降低了37.77%和67.06%、15.68%和30.62%、28.72%和69.09%;籽粒蛋白质含量显著提高了4.33%和10.16%,籽粒淀粉含量显著降低了5.65%和22.78%,籽粒饱满度显著降低了12.83%和18.95%。在同一胁迫期内,除了籽粒蛋白质含量以外,其余各项测定指标值均随着复水量的减少而不同程度的降低,指标测定值均表现为W1W2W3,且在T2和T3胁迫期下较对照W1,W3降低尤为明显。通过测定饲草大麦叶片叶绿素含量发现:在T1和T2持续胁迫时间下,复水后W1和W2处理存在显著的补偿效应,而W3以及T3持续胁迫时间的所有复水处理均未产生显著的补偿效应。说明甘饲麦1号干旱复水补偿效应的利用应注意干旱胁迫持续时间不超过20 d(土壤最大持水量的60%～65%),旱后复水量不低于田间土壤最大持水量的55%～60%为宜。     

柑橘脉突病毒基因组全长cDNA克隆及其侵染性鉴定

【目的】构建柑橘脉突病毒（citrus vein enation virus,CVEV）侵染性克隆,为从分子水平解析其致病机理打下基础。【方法】利用SMARTer^? RACE(rapid amplification of cDNA ends)试剂盒对CVEV的5′序列进行RACE,并依据序列分析结果及CVEV分离株VE-1保守序列,设计CVEV基因组全长cDNA扩增引物。以CVEV毒源植株的总RNA为模板,通过EV25-F/EV5983-R引物扩增CVEV基因组全长cDNA。利用In-Fusion重组连接线性化pXT1和CVEV全长cDNA。通过菌液PCR及测序分析鉴定CVEV基因组全长cDNA克隆。通过农杆菌介导的真空浸润接种摩洛哥酸橙（Citrus aurantium）、邓肯葡萄柚（C. paradisi）、尤力克柠檬（C.limon）、枳柚（C. paradisi×Poncirus trifoliata)、Rusk枳橙（P. trifoliata×C. sinensis）、枣阳小叶枳(P. trifoliata),进一步通过RT-PCR检测、症状观察鉴定所构建CVEV全长cDNA克隆的侵染性。【结果】建立了CVEV的基因组全长RT-PCR扩增体系,获得基于双元载体pXT1的CVEV基因组全长cDNA克隆10个。随机选取的6个全长cDNA克隆CVEV1901—CVEV1906的序列一致性为99.35%。其中,CVEV1901基因组全长5 983 nt,由5个开放阅读框、5′端207 nt和3′端198 nt的两个非翻译区、以及ORF2和ORF3之间122 nt的基因间隔区组成。序列分析结果显示,CVEV1901与浙江分离株XZG及四川SM分离株的序列一致性分别为99.98%和99.11%;与西班牙VE-1分离株、美国加州VE701分离株和日本IBK分离株基因组序列一致性在96.89%—98.61%;与同属中豌豆耳突花叶病毒（pea enation mosaic virus）和紫花苜蓿耳突病毒（alfalfa enamovirus）的序列一致性约90%。通过农杆菌介导的真空浸润将CVEV1901接种至6个不同的柑橘品种,接种后120 d的RT-PCR检测结果表明摩洛哥酸橙、邓肯葡萄柚、尤力克柠檬、枳柚、Rusk枳橙和枣阳小叶枳阳性植株/接种植株（阳性率）分别为16/17（94.12%）、12/14（85.71%）、16/21（76.19%）、15/19（78.95%）、13/14（92.86%）和0/18（0）。其中,部分摩洛哥酸橙出现典型CVEV侵染症状,叶片侧脉和支脉产生耳状小突起,叶背有相应的凹陷;部分邓肯葡萄柚和尤力克柠檬出现叶片皱缩现象。【结论】建立了CVEV的基因组全长RT-PCR扩增体系,获得了CVEV基因组全长cDNA侵染性克隆,通过农杆菌介导的真空浸润接种可引起摩洛哥酸橙、邓肯葡萄柚和尤力克柠檬的CVEV侵染症状。     

避雨栽培对‘红阳’猕猴桃光合作用及果实品质的影响

以露地和避雨栽培条件下的‘红阳’猕猴桃为试验材料,通过测定园地温湿度、植株光合作用、果实品质等指标,研究避雨栽培对‘红阳’猕猴桃生长微环境、叶片光合能力和果实品质的影响。结果表明:避雨栽培条件下植株环境温度降低2.4℃,空气湿度提高12.2%,光合有效辐射下降13.0%~28.6%;避雨栽培可维持‘红阳’猕猴桃在生长旺盛期的平均净光合速率,有效缓解植株光合"午休"现象,对植株有一定的保护作用,表现为结果枝数降低10.8%,结果枝长度增加60.8%,坐果数提高129.7%,叶绿素含量提高6.1%,单果重提高25.7%,采收期和软熟期可溶性固形物含量分别增加10.8%和14.1%,干物质含量无显著差异,延长了贮藏时间。综上,避雨栽培可改善‘红阳’猕猴桃生长微环境,优化植株光合作用,提高果实品质。     

SRT1720 enhances maturity and quality of oocytes in aged mice

This study aims to investigate the morphology and distribution of mitochondria, spindles, and chromosomes in oocytes of aged mice and examine the effects of SRT1720 on oocyte maturation. C57BL/6J mice were divided into young (4–8 weeks) and aged groups (48–52 weeks). In vitro maturation media contained (0.05, 0.1, and 1.0 μM) SRT1720 and 0.1-μM dimethyl sulfoxide (DMSO control). The rate of chromosome misalignment and spindle misorientation in oocytes of aged mice were significantly higher than that of young mice (P < 0.01). Fluorescence intensity of mitochondria from oocytes of aged mice was significantly lower than that of young mice (P < 0.01). SRT1720 at 0.1 μM significantly improved oocyte maturation, fertilization, and blastocyst formation in aged mice compared with young mice (P < 0.01). Additionally, immunofluorescence intensity of mitochondria, normal spindle morphology, and chromosome alignment were notably enhanced with SRT1720 when compared with the DSMO control group for metaphase II (MII)-stage oocytes matured in vitro (P < 0.01); 0.1-μM SRT1720 enhanced the expression level of SRIT1 in oocytes from aged mice. In summary, the aged mice oocytes showed increased nuclear and cytoplasmic defects, whereas SRT1720 enhanced oocyte maturation and quality. We concluded that 0.1-μM SRT1720 was an appropriate concentration for in vitro maturation media.     

雄性食蟹猴生殖系统中Ghrelin的分布定位

本文采用免疫组织化学染色方法,探讨ghrelin在雄性食蟹猴生殖系统内的分布定位。通过对ghrelin免疫阳性细胞在生殖系统中分布部位、含量及细胞形态等方面的研究,为今后ghrelin在食蟹猴体内的功能研究奠定形态学基础。免疫组化染色发现,ghrelin阳性细胞在食蟹猴的生殖系统中有分布。Ghrelin免疫阳性细胞被染为棕色到棕黑色,主要分布于睾丸、附睾及输精管中,精囊腺中无ghrelin阳性细胞分布。Ghrelin阳性细胞在组织中多呈散在分布。细胞大小不一、形态各异,多呈圆形、卵圆形、锥体形、长柱形及其他不规则形。     

Pyroptosis mediates high glucose-induced inflammation and injury in MC3T3-E1 osteoblasts

AIM To investigate whether pyroptosis contributes to the inflammation and injury in mouse embryonic osteoblastic cell line MC3T3-E1 induced by high glucose (HG; 45 mmol/L glucose). METHODS The cell viability was measured by CCK-8 assay. The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 (CASP1) were determined by Western blot. The secretion levels of interleukin-18 (IL-18) and IL-1β were measured by ELISA. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The alkaline phosphatase (ALP) activity was determined using the ALP kit, and the number of mineralized nodules was detected by alizarin red S staining. RESULTS After the MC3T3-E1 osteoblasts were treated with HG for 24 h, the protein expression levels of NLRP3 and CASP1, and the secretion levels of IL-18 and IL-1β were significantly increased. The decrease in cell viability, and the increases in ROS generation and MMP loss were also observed. Moreover, the differentiation and mineralization of MC3T3-E1 osteoblasts were inhibited, evidenced by decreases in both ALP activity and mineralized nodule number. Knockdown of CASP1 by siRNA attenuated the HG-induced osteoblast inflammation and injury mentioned above. CONCLUSION Pyroptosis mediates HG-induced inflammation and injury in MC3T3-E1 osteoblasts.     

Effects of different active constituents of Gynostemma pentaphyllum on mitochondrial energy metabolism-related proteins in endothelial cells induced by ox-LDL

AIM To investigate the effects of different components of Gynostemma pentaphyllum ［gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)］ on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a （Cox5a）, NADH:ubiquinone oxidoreductase core subunit S1 （Ndufs1）, ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C）, were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P<0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P<0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P<0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P<0.05 or P<0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis.